CN106662530A - Method and apparatus for optical measurement of liquid sample - Google Patents
Method and apparatus for optical measurement of liquid sample Download PDFInfo
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- CN106662530A CN106662530A CN201580047404.9A CN201580047404A CN106662530A CN 106662530 A CN106662530 A CN 106662530A CN 201580047404 A CN201580047404 A CN 201580047404A CN 106662530 A CN106662530 A CN 106662530A
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/75—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated
- G01N21/76—Chemiluminescence; Bioluminescence
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/01—Arrangements or apparatus for facilitating the optical investigation
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/6408—Fluorescence; Phosphorescence with measurement of decay time, time resolved fluorescence
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2201/00—Features of devices classified in G01N21/00
- G01N2201/06—Illumination; Optics
- G01N2201/062—LED's
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Abstract
A method and an apparatus for optical measurement of a liquid sample (2) placed in a sample well (3) where the sample in the sample well is exposed to excitation light (7) at first wavelength from an excitation light source (4), which excitation light generates a collection of singlet state oxygen molecules from donor molecules in the liquid sample, said singlet state oxygen molecules reacting with acceptor molecules in the liquid sample causing said acceptor molecules to emit chemiluminescence emission light (11) at second wavelength, which second wavelength is shorter than said first wavelength, and where the emission light produced by the excitation light is measured with a detector, characterized in that the excitation light source is a LED (4).
Description
Technical field
The present invention relates to the optical measurement of fluid sample.More precisely, the invention relate to be placed in sample well
Fluid sample optical measurement method and apparatus, the wherein sample in sample well is exposed to exciting from excitation source
Light, and wherein launched by the light of generation that excites of light with detectors measure.
Background technology
Fluorescence is the light carried out by the material for absorbing light or other electromagnetic radiation（Photon）Transmitting.The absorption of energy will
The orbital electron of molecule is energized into higher electronic state and relaxes towards ground state emission and goes out photon.This is a kind of luminous
（luminescence）Form.Generally, compared to the radiation for being absorbed, the light launched have longer wavelength, and therefore
With relatively low energy.
Chemiluminescence is due to light transmitting caused by chemical reaction（It is luminous）.
Fluorogen absorbs the luminous energy at a wavelength, and as response, re-emits the light at another longer wavelength
Energy.Each fluorogen has a light absorbing distinctive wave-length coverage in this place, and another of launching light in this place has area
Other wave-length coverage.The property allows them to be used for the particular detection of biological products by analytical instrument and technology.
From the different sample measuring methods based on luminescence generated by light known in the art, wherein swashing using the light into sample
Send out the light transmitting obtained from sample.When the light by electromagnetic radiation is measured, it may be determined that the heterogeneity of sample.Known measurement
Method includes the AlphaScreen described in the United States Patent (USP) 6,409,913 of Ullman et al.（Amplify luminescent proximity uniformly to determine
Screen）Photochemistry e measurement technology and LOCI（Luminescent oxygen passage immunoassays）, wherein exciting and chemical reaction two in sample by light
Person produces light transmitting.
The light source being usually used in AlphaScreen e measurement technology method and apparatus is laser, due to the ripple of the light of generation
The high power density of long uniformity and the light of acquisition, the high excitation energy effectively causes from the pearl for being coated with luminous agent and is formed
The oxygen molecule of modification.The final initiating chamical reaction cascade of oxygen molecule of modification, causes the generation of detected chemiluminescence signal.
AlphaScreen measuring methods are that based on the neighbouring measure of pearl, wherein donor bead and acceptor bead is by can specificity
React to each other and with pearl reaction various biological analyte materials be connected.When the red laser with 670-690nm wavelength swashs
When sending out the donor bead of connection, periphery oxygen is converted into excited singlet state by the donor bead of the connection.The creating singlet oxygen of release is diffused into
The sample well matrix of surrounding, and if donor bead and acceptor bead are via particular organisms reaction forming to be determined, acceptor bead foot
It is enough close to be excited by singlet state, and light is transmitted in 520-620nm wavelength（It is blue-green, green or gold-tinted）Between produce and
It is proportional to interaction level.By measuring launching light, it is possible to establish significant degree that chemical reaction occurred and
What property of the sample of measure.
Wavelength difference between the exciting light and launching light of sample allows to separate and detect the light from excitation light emission.
In AlphaScreen methods, it is however generally that the wavelength of exciting light and launching light is compared with fluorescence with contrary
Relation, i.e. excitation wavelength are longer than wavelength of transmitted light.
In fluorescence measurement, generally, excitation light pulse is sent to sample, and be effectively realized the measurement of launching light, and
Delay is not induced.In time resolution fluorescence spectral（TRF）In, after one or more excitation light pulses, according to the time
Function monitors sample.Background fluorescence is reduced using TRF, and for example in dynamics research.
Laser is potentially a kind of accurate and powerful light source for generally producing collimated light beam, and the collimated light beam is reduced
The needs of single collimator apparatus used in analytical instrument, but also produce some problems using laser as excitation source.Produce
High electrical power required for raw laser also generates heat, but is because the hot and temperature that the resonator of laser diode is generated to oneself
Change is extremely sensitive, so laser diode needs the regular active of the such as specific installation of peltier cooler etc
Cooling, this causes to increase the complexity and size of analytical equipment.Further, laser diode is also very expensive, and this improves
Using the price of the analytical equipment of laser instrument.In practice it has been observed that when with such as swashing in AlphaScreen applications
During light source activation sample, only some measurements, typically only one of which measurement can be performed, because in follow-up measurement, obtaining
Obtain significantly reduced transmission signal or without transmission signal.
By using appropriate stable fluorescence reference material（Shorter wavelength exciting radiation is converted into longer wavelength by it
Transmitting radiation）Fluorescence can be stablized and read instrument.The material of these species is broadly available and for example for Thermo
The Fluoroskan Ascent microplate fluorometers of Scientific.
In Alphascreen measurements, excitation wavelength（685nm）Compare launch wavelength（520nm-620nm）It is long.Do not exist by
The excitation wavelength of 685nm is converted into the available appropriate stabilizing material of shorter wavelength.This is problematic, because to compared with shortwave
One of benefit that long shift is in fact measured for Alphascreen, because there is no optical background signal in the measurements.Equally,
The reference material for using is showed in default of TRF（behavior）, it is impossible to survey in the TRF including Alphascreen chemical processes
It is used as to refer to fluorescent material in amount.
The content of the invention
The present invention provides a kind of novel method and apparatus of the optical measurement for fluid sample, when exciting photogenerated list
Weight state oxygen（It further causes to launch chemiluminescent light）When so that the wavelength of exciting light is more than the chemiluminescence emission observed
The wavelength of light.
The method and device can be used for the optical measurement of fluid sample, for example for using fluorescence Alphascreen or its
The chemical reaction that he is determined based on creating singlet oxygen passage, such as in AlphaScreen, AlphaLISA and similar optical measurement
In method.The creating singlet oxygen passage is determined and may imply that based on by chemiluminescent measure caused by creating singlet oxygen, wherein exciting
The wavelength of the chemiluminescence emission light that optical wavelength ratio is observed is longer.
Excitation source can be LED（Light emitting diode）.In the case of using LED, can obtain and be swashed using semiconductor
Light device identical luminous power, without the quality by exciting light caused by temperature change（In view of laser diode can be subject to even
The impact of single degree Celsius of temperature change）Problem.The use of LED also allows the structure of measuring apparatus compacter.
However, LED makees used in the chemical reaction application determined in Alphascreen or based on other creating singlet oxygen passages
Major advantage for excitation source is that measured launching for sample will not be to subtract with the measurement identical mode using laser
It is few, it means that identical sample is in different time points（Dynamics research）Can be excited and measure several times（At least twice）.
Kinetic measurement is generally used for studying various enzymatic reactions.The example of dynamic experiment is described in example 1 and Fig. 5 A to Fig. 5 C,
Wherein during the process of chemical reaction, identical sample is measured repeatedly in each setting time point, to monitor reactive chemistry
Dynamics.
In the present invention, after a suitable period of time, using single exposure and single transmitting reading can be excited to carry out
TRF is measured, and such as 300ms exposures, 50ms postpone and 500ms transmittings are read.Alternately, in desired time of measuring, profit
Practicable TRF measurements are circulated with multiple continuous measurements, wherein being short emission measurement after short excitation pulse, for example, is swashed by 5ms
It is that 5ms transmittings are read the 10ms of composition and excite and launch reading to circulate in be repeated continuously in desired time of measuring after sending out.
When the amount of background is reduced, instrument becomes more sensitive, and can more reliably detect relatively low analyte concentration.
The problem of background products can in many ways be reduced.Instrument hardware may be designed as reducing background but for example wave filter leads to
It is not often 100% accurate, because some exciting lights may pass through emission filter, vice versa.After a certain period of time, background
Transmitting exhausts.While transmitting is read, TRF exciting lights are not illuminated, and when reading, most of background emissions are also
It is decreased.Equally in Alphascreen or other technologies based on creating singlet oxygen passage measure, launch wavelength is less than excitation wave
It is long, it means that not to be read at electromagnetic radiation identical wavelength more than the background fluorescence signal of excitation wavelength.Using confocal
Optical devices reduce the amount of the background signal of generation also by the irradiation in the other parts minimized outside sample liquids.
The present invention it is a kind of for being placed on sample well in fluid sample optical measurement method in, in sample well
Sample be exposed to exciting light from excitation source first wave strong point, donor molecule of the exciting light from fluid sample is generated
The set of singlet state molecular oxygen, the singlet state molecular oxygen is reacted so as to cause the acceptor with the acceptor molecule in fluid sample
Molecule launches chemiluminescence emission light with second wave length, and second wave length is surveyed with detector here than the first wave length
The launching light produced by exciting light is measured, wherein excitation source is LED.
In one embodiment of the inventive method, the open area of sample well is focused on from the exciting light of LED, and
And collect launching light from the open area of sample well using confocal arrangement.Present embodiments provide effective and uniform photograph of sample
Penetrate, and extremely sensitive measurement result.
In the context of the present invention, the use of term " confocal " means the focus of exciting light and Jiao of launching light set
Point actually one, and the same focal point of arrangement and the shared at least one common optics unit of the optical path of launching light
Part, such as lens, speculum or beam splitter.
In one embodiment of the inventive method, exciting light is imported into sample well, and the sample from confocal arrangement
Well collects launching light, and the image of wherein excitation source is focused in the open top of sample well, and the light-receiving of detector
The image in region is focused in the open top of sample well.In this embodiment, the described image of excitation source is smaller than sample
The opening of product well, and the described image of the optical receiving region of detector can be more than the opening of sample well.
In one embodiment of the inventive method, to sample excitation light pulse, and sending out from excitation light pulse are sent
Send and pass by after predetermined time period, carry out the measurement of launching light.Predetermined time period can be 30-70ms, for example, lead to
It is often 50ms.
In one embodiment of the inventive method, exciting light and launching light are calibrated.Preferably by separately and independently
Calibration process is carrying out the calibration.
In one embodiment of the inventive method, LED used is in 680nm emitted at wavelengths light.
In one embodiment of the inventive method, the inwall of sample well is formed by diffuse-reflective material or is coated with diffusing reflection
Material.This inwall of well is that both exciting light and launching light provide good diffusing reflection surface, and this improves measurement process
Quality.The example of suitable material is the polystyrene with white colour material, such as titanium oxide.
The present invention it is a kind of for being placed on sample well in the device of optical measurement of fluid sample include for the
To the excitation source of the sample in exciting light exposed sample well at one wavelength, and at second wave length from sample detection simultaneously
The detector of measurement chemiluminescence emission light, the second wave length is less than the first wave length, and wherein excitation source is LED.
In one embodiment of apparatus of the present invention, the device is included for exciting in the open area for focusing on sample well
The focus set of the image of light source, and the figure of the optical receiving region for the detector in the open area for focusing on sample well
The equipment of picture, here confocal point is actually in the level of the opening of sample well.
In one embodiment of apparatus of the present invention, excitation source is pulse excitation light source.
In one embodiment of apparatus of the present invention, the device also includes exciting optical filter, transmitting optical filter, excites
With transmitting beam splitter and the spuious light shield above sample well.
In one embodiment of apparatus of the present invention, the device also includes exciting beam splitter and for calibrating exciting light
Excite reference detector, and/or transmitting reference light source and the white reflective scattering surface for calibrating emission detector.
In one embodiment of apparatus of the present invention, LED is excited for the LED emission light at 680nm wavelength.
In one embodiment of apparatus of the present invention, detector is photomultiplier.
The feature for limiting the inventive method is stated more precisely in claim 1, and limits the feature of apparatus of the present invention
It is stated more precisely in claim 17.Dependent claims disclose the favorable characteristics and embodiment of the present invention.
Description of the drawings
In the following example, some embodiments are more fully described with reference to the accompanying drawings, wherein
Fig. 1 schematically illustrates by way of example measurement arrangement of the invention,
Fig. 2A and Fig. 2 B to be schematically illustrated excite and launch pattern using what the present invention was obtained by way of example,
Fig. 3 schematically illustrates by way of example the image used in the present invention,
Fig. 4 schematically illustrates by way of example the improvement embodiment that arrangement is measured in Fig. 1, and
Fig. 5 A to Fig. 5 C show the result of the kinetic measurement carried out using instrument of the invention.
Specific embodiment
Schematically illustrate in Fig. 1 for realizing the appropriate e measurement technology application determined based on creating singlet oxygen passage（It is all
Such as such as AlphaScreen or AlphaLISA）Measurement arrangement be included in the optics of fluid sample 2 in sample well 3 and survey
The optical measuring apparatus 1 of the present invention during amount, the sample well for example can be a part for 384 microwell plates.
Optical measuring apparatus 1 include the LED4 of the excitation source that equipment is formed using lens arrangement 5 and wave filter 6.LED4
In 680nm emitted at wavelengths light, the light is collected into light beam with lens arrangement 5 and using wave filter 6 it is filtered to protect
The appropriate wavelength of the excitation beam 7 that card is achieved in that.
Then the beam splitter 8 of reflected light at 680nm wavelength is used in towards the reflected excitation light beam 7 of sample well 3.In excitation beam
7 leave measuring apparatus 1 is before focused on excitation beam 7 using lens arrangement 9 so that the focus of excitation beam is located at sample well 3
Open area in.
When excitation beam 7 is focused in the open area of sample well 3, the inner surface of the sample well 3 is white, the sample
Product well is shown as Ulbricht（Ulbricht）Integrating sphere is the same, and effectively lights the whole liquid inside sample well
Sample 2.This point is more closely discussed later in reference to Fig. 2A and Fig. 2 B.
Profit is collected from the confocal focus of identical rather than where excitation beam is focused in the open area of sample well 3
The launching light obtained from fluid sample 2 with excitation beam 7.Crosstalk shields thing 10 with aperture is located adjacent to opening for sample well 3
Above mouthful, this passes through possible exciting light of the optics minimum stray light from adjacent well.
The launching light collected from fluid sample 2 is directed to the generally transmitting light beam 11 with 520nm to 620nm wavelength.
These wavelength are not reflected and through beam splitter 8 by beam splitter 8.Beam splitter 8 can also be the broadband mirrors of part reflection, this
It is useful when needing free wavelength to select.Filtered with 12 pairs of transmitting light beams 11 of wave filter and will launch light beam 11 with lens 13
Focus on the detection surface of detector elements 14, the detector elements 14 are in the present embodiment the negative electrode of photomultiplier.
Using the measuring apparatus 1 of Fig. 1, exciting beam 7 can be generated and the collection with transmission signal beam 11 and measurement simultaneously
Send exciting beam 7（It is attributed to beam splitter 8）, it is preferable that excitation beam is generated as one or more pulses, and from
Transmission pulse was pass by after the predetermined time, carried out the collection and measurement of transmission signal（TRF is measured）.
Fig. 2A and Fig. 2 B to be schematically illustrated and excite and launch pattern using what the present invention was obtained.Fig. 2A is shown according to bright
Excite the reception pattern, and Fig. 2 B of primary cosine law shows the radiation diagrams case according to Lambert's cosine law.
White micropore 3 or the micropore with white inner surfaces are shown as Ulbricht integrating sphere.Ulbricht
Opening on integrating sphere is shown as preferable diffusing radiation surface, and thus follows Lambert's cosine law.Radiation pattern
For the circle in the open top of well 3.Radiate when receiving（Excite）With transmission radiation（Transmitting）When, opening shows as identical.
Just it is placed on the opening of well 3 by the way that confocal point will be excited, maximum solid angle can be used to excite, and by inciting somebody to action
Launch confocal point to be placed on the opening of well, maximum solid angle can be used for transmitting and read.
When collimated light beam to be fitted to the opening of well 3, the very short Optical devices of focal length can be used.This means LED
The amplification of output can remain it is low, and the LED focusedimages at plate opening less than opening size.This causes high exciting
Efficiency.Further, because the LED image on opening is less than the size of opening, the position of well is not crucial.
Exciting beam is completely smooth in the inside of well 3, and for well inside gentle irradiation is provided.Particle and liquid component from
Above, below and side are uniformly illuminated.This minimize the risk of many bleaching effects.
In transmitting is read, using the very short lens of same focal length, so that the opening for envisioning well 3 meets by interfering filter
First collimated light beam of ripple device, and and then transmitting beam be focused on the negative electrode of photomultiplier.Short focal length lens can be with
Very big angular collection carrys out the light of the opening of artesian well 3, this generates high transmitting collection efficiency.Also, due to the figure of well opening
As the negative electrode less than photomultiplier, the positioning of well is not crucial.
Exciting for optics minimum stray light adjacent well can be passed through using other light shield.Identical light shield is also most
The little transmitting crosstalk changed during transmitting is read.For example, the spuious light shield of combination can be 1：10000.
Fig. 3 schematically illustrates the image that the present invention is used.Show exciting light LED4 on the opening of well 3 in Fig. 3
Image 15, the well is a part for the well plate for including multiple wells 3.Compared with the open area of well 3, the negative electrode 14 of photomultiplier
Image 16 it is very big.Using crosstalk shields thing 10（Opening of its opening 17 less times greater than well 3）Adjacent well is effectively prevented to the moon
The compromising emanation of pole 14.
Fig. 4 shows the improvement embodiment that arrangement is measured in Fig. 1, and wherein optical measuring apparatus 1 ' are equipped with for exciting light
With the calibration system of both launching lights.
Instrument reads response for launching efficiency and the product of transmitting response.Therefore, in the improvement embodiment of the present invention,
By individually calibrate launching efficiency and transmitting response can stable optical measuring apparatus 1 ', so as to calibrate be divided into two it is independent
Calibration process：Excite calibration and transmitting calibration.
Launching efficiency is calibrated relative to short-term and change in long term.
Short term variations are present in excitation light pulse launched by power LED 4, usually 100ms to 500ms length
Portion.The LED of these species is not heat-staple in the power level for needing, therefore excitation light pulse amplitude can be in the burst length
Period changes.This can change the accurate clocking effect of the LED excitation pulses of generation.Excitation pulse shape（I.e. wave-length coverage and
The light quantity produced at each wavelength）Must be repeatable to preserve accurate timing, and it is sharp to measure the average or peak value of pulse
Luminous power is inadequate.LED output levels must be continuously controlled and adjust within the excitation pulse time, to remain permanent
Fixed effective light output.
In the fig. 4 embodiment, beam splitter 18 is located in excitation light path, and it is little to exciting reference detector 19 to separate one
Partial exciting beam 7.By the electric current for controlling and being adjusted to LED4, the output signal for carrying out self-excitation reference detector 19 is protected
Hold constant.Control circuit can be used（It is not shown）Realize control and be adjusted to the electric current of LED4, the control circuit includes proportional integral
Controller（PI- controllers）With the power stage of the electric current for being generated to LED.This excites calibration system to set in the optical measurement of the present invention
The excitation light pulse of precise constant amplitude is generated in standby 1 '.
This excites calibration system and also provides for well to well and plate to the exciting light that plate is measured and long-term excites stability.
In transmitting calibration, photomultiplier 20 is referred to as drift with the exporting change of operating time, and these float
Shifting is divided into short term drift and long term drift.
By using the stable calibration of transmitting reference light source 21, these drift about.Stable transmitting reference light source 21 can be
LED, such as temperature especially stable Philip LXML-PR00-0500.The wavelength of the light of LED emission is 460nm.These species
LED for the purpose be excessively powerful transmitter, and output should be strongly attenuated（For example utilize neutral density
Wave filter）.
In the fig. 4 embodiment, emission filter 12 is rotation emission filter wheel, wherein existing for eight transmitting filters
The position of ripple device, not covering for the hole of measurement and for launching background signal measurement with wave filter（shutter）Behaviour
Make solid section.In hidden position, there is the white reflective scattering surface for being launched the irradiation of reference light source 21.In transmitting calibration
Period, emission filter wheel is rotated in hidden position, and the white reflective scattering surface transmitting on emission filter wheel
Reference light source 21 irradiates, and the reflected light being scattered equably irradiates the negative electrode 14 of photomultiplier 20, and measures calibration water
It is flat.
For transmitting calibration, original photomultiplier calibrated horizontal can be stored in optical measuring apparatus 1 ' during its manufacture
In.Transmitting calibration for example can be carried out before each minitype plate reads, and by comparing calibration result and the storage of measurement
Original photomultiplier calibrated horizontal value calculates correction factor.
The final response of system is the product of booster dose and transmission signal.This is especially particularly important in TRF measurements.When
Light source（Transmitting）Intensity when being arranged to constant, measure booster dose（And need not other corrections）Be enough to define system sound
Should.
Device 1 can be arranged to form LED4 focusedimages at the top of the opening of sample well 3 by focusing on the light of LED4
15.Device 1 may include lens arrangement 9, provide narrow beam by focusing on the light of LED so that in the focal point of narrow beam
Form focusedimage 15.The numerical aperture of lens arrangement 9（NA）0.15 can be for example more than, more than 0.20, more than 0.30, be more than
0.40, more than 0.50 or even greater than 0.80.Using big solid angle and open top focusedimage can for sample well 3 in contain
Some samples 2 are provided effectively and uniformly irradiated.By using diffusing reflection sample well 3 and big solid angle, device 1 can be arranged
Into from above, below with side equably irradiating sample 2.Uniform irradiation can reduce the risk of photobleaching effect.
Because positioning is inaccurate, the opening of sample well 3 can be relative to focus somewhat（Flatly）Dislocation.Uniform irradiation can subtract
The effect of the little horizontal location mistake with regard to measurement result.
The upright position of the upper surface of the fluid sample contained in sample well 3 can also have the effect with regard to measurement result.
In fact, the physical location of upper surface can be slightly offset from reference position.Uniform irradiation can reduce with regard to the described inclined of measurement result
Poor effect.
LED can be powered by the electric current coupled to LED.The output intensity of LED and the ratio of electric current can near the operating point of LED
Being substantially linear.Supplied by using electric current, can easily control LED.Therefore, the electric current of LED is coupled to by control
The output intensity of LED can easily be stablized.By reducing the short term variations inside single excitation light pulse, can further reduce light
The risk of bleaching effect.Device 1 can be arranged to the substantially invariable effective light output of maintenance during single excitation light pulse.It is logical
The instantaneous strength of the intensity controlled excitation light pulse to monitor excitation light pulse is crossed using reference detector 19 is excited.The method
May include the electric current for adjusting LED4 so that the output signal for carrying out self-excitation reference detector 19 is kept during excitation light pulse
For substantially invariable.For example by using based on the pi controller from the output signal for exciting reference detector 19 to obtain
（PI- controllers）The electric current of LED4 can be generated.A part of light of excitation light pulse can be optically coupled to reference detector 19, so as to
Output signal is provided.The part can be optically coupled to reference detector 19, such as by using beam splitter 18.
LED4 can have with the accurate light-emitting zone limited with stable dimensions.Intensity distribution can on the surface of light-emitting zone
Being spatially substantially homogeneous.The light-emitting zone of LED4 can have substantially flat output surface.The image of light-emitting zone
Can be focused in the open top of sample well 3.Light-emitting zone can have stable length and width so that the figure of light-emitting zone
The positions and dimensions of picture can also be extremely stable.LED can have generally rectangular light-emitting zone or almost circular luminous
Region, and sample well 3 can have generally rectangular or circular opening.The shape of optional light-emitting zone, to match sample
The opening shape of well 3（Or vice versa it is as the same）.Therefore, the picture shape in rectangle or circular luminous region can match opening for sample well 3
Mouth-shaped.In one embodiment, the sample well 3 of sample panel can be arranged with rectangular array so that the well of plate has basic
The opening of rectangle, and the image of rectangular emitting area can be focused in the open top of the well 3 of the well plate.
LED can provide incoherent exciting light 7, and this can reduce the risk of photobleaching effect.
Thanks to the uniform controlled irradiation by using LED, photobleaching is in the chemiluminescent light from electromagnetic radiation
Intensity aspect can have the effect for reducing.Sample can be excited by several continuous excitation light pulses so that described continuously to excite
The chemiluminescent luminous intensity detected after the last exciting light of light pulse still can be substantially greater than zero.Especially, exciting
Combination degree during light pulse in sample is swashed in the case of holding identical, and sample can swash by the first excitation light pulse and by second
LED pulse is excited so that the intensity of the chemiluminescent light launched after the second excitation light pulse is more than in the first exciting light arteries and veins
80% or or even the 90% of the intensity of the chemiluminescent light launched after punching.
The combination of the molecule captured on pearl can have on the energy transmission from creating singlet oxygen to acceptor bead to be affected.Thus,
The intensity and/or energy of chemiluminescence emission light may indicate that the combination degree in sample.Thanks to the photobleaching for reducing, by exciting
The amount of the singlet state molecular oxygen that light pulse is generated can remain substantially invariable or be reduced with very slow speed.This can be allowed in sample
Combination degree is more accurately monitored.
The method may include：
- with the first excitation light pulse irradiating sample 2, to generate the first collection of singlet state molecular oxygen from the donor molecule of sample 2
Close,
- cause the first chemiluminescence to send out to the acceptor molecule transmission energy of sample 2 by the first set from singlet state molecular oxygen
The transmitting of light is penetrated,
The first chemiluminescence emission light of-measurement,
- with the second excitation light pulse irradiation identical sample 2, to generate the of singlet state molecular oxygen from the donor molecule of sample
Two set,
- cause the second chemiluminescence to send out to the acceptor molecule transmission energy of sample 2 by the second set from singlet state molecular oxygen
The transmitting of light is penetrated, and
The second chemiluminescence emission light of-measurement,
Wherein, sample 2 is by the first excitation light pulse and by the irradiation of the second excitation light pulse so that generated by the second excitation light pulse
Singlet state molecular oxygen amount the singlet state molecular oxygen generated by the first excitation light pulse amount 80% to 100% scope
It is interior.
The method may include：
- with the first excitation light pulse irradiating sample 2, to generate the first collection of singlet state molecular oxygen from the donor molecule of sample 2
Close,
- cause the first chemiluminescence to send out to the acceptor molecule transmission energy of sample 2 by the first set from singlet state molecular oxygen
The transmitting of light is penetrated,
The first chemiluminescence emission light of-measurement,
- with the second excitation light pulse irradiation identical sample 2, to generate the of singlet state molecular oxygen from the donor molecule of sample
Two set,
- cause the second chemiluminescence to send out to the acceptor molecule transmission energy of sample 2 by the second set from singlet state molecular oxygen
The transmitting of light is penetrated,
The second chemiluminescence emission light of-measurement,
Wherein sample 2 is by the first excitation light pulse and by the irradiation of the second excitation light pulse so that the second chemiluminescence emission light
Maximum intensity is more than or equal to the 80% of the maximum intensity of the first chemiluminescence emission light, and the combination degree wherein in sample
Keep identical during the first excitation light pulse and the second excitation light pulse.
The method may include：
- with the first excitation light pulse irradiating sample 2, to generate the first collection of singlet state molecular oxygen from the donor molecule of sample 2
Close,
- cause the first chemiluminescence to send out to the acceptor molecule transmission energy of sample 2 by the first set from singlet state molecular oxygen
The transmitting of light is penetrated,
The first chemiluminescence emission light of-measurement,
- with the second excitation light pulse irradiation identical sample 2, to generate the of singlet state molecular oxygen from the donor molecule of sample
Two set,
- cause the second chemiluminescence to send out to the acceptor molecule transmission energy of sample 2 by the second set from singlet state molecular oxygen
The transmitting of light is penetrated,
The second chemiluminescence emission light of-measurement,
Wherein sample 2 is by the first excitation light pulse and by the irradiation of the second excitation light pulse so that the second chemiluminescence emission light
Maximum intensity is more than or equal to the 90% of the maximum intensity of the first chemiluminescence emission light, and the combination degree wherein in sample
Keep identical during the first excitation light pulse and the second excitation light pulse.
The maximum intensity of the first excitation light pulse is substantially equal to the maximum intensity of the second excitation light pulse, and first
The energy of excitation light pulse is substantially equal to the energy of the second excitation light pulse.From the intensity of the excitation light pulse of LED emission
Substantially constant can be also kept during excitation light pulse.The duration of optional light pulse so that the second excitation light pulse is led
The maximum intensity of the chemiluminescence emission light of cause is for example higher than ten times of the minimum detection limit of device 1.May be selected first to excite
The intensity of the intensity of light pulse and the second excitation light pulse so that by chemiluminescence emission light caused by the second excitation light pulse
Maximum intensity is higher than 10 times of the minimum detection limit of such as device 1.The minimum detection limit may imply that can be with ambient noise
The minimum strength of the transmitting mutually distinguished.When intensity is for example higher than ten times of the minimum detection limit, can reasonably precision measure
The value of intensity.
It is measurable to be excited by first during detection time section between the first excitation light pulse and the second excitation light pulse
The intensity of exciting light caused by light pulse.Time delay between first excitation light pulse terminates and the second excitation light pulse starts
30ms can be for example more than or equal to.Selecting time postpones that the second excitation light pulse is caused in the first excitation light pulse of measurement
Chemiluminescence emission used by detection time section terminate before will not start.The intensity of exciting light is in the detection time section phase
Between be substantially equal to zero.Time delay can for example be more than 30ms, more than 100ms, more than 500ms, more than 1s, more than 10s, greatly
In 60s, more than 10min, more than 1h, more than 2h, more than 12h or even greater than 24 hours.Can be with the first exciting light arteries and veins
Rush after irradiating sample with the second excitation light pulse irradiating sample 2.The maximum intensity of the first excitation light pulse is substantially equal to
The maximum intensity of the second excitation light pulse.Alternatively, the signal for obtaining from detector can be integrated, and detection time section
Also referred to as integration time period.
Obtaining chemiluminescence emission light several times from same sample can provide more reliable and/or more accurate measurement result.From
Same sample obtains several times chemiluminescence emission light can promote the analysis of Time Dependent phenomenon.
In one embodiment, can be with least ten continuous excitation pulses equably irradiating sample so that chemiluminescence
The intensity of launching light is even substantially greater than zero after the tenth excitation light pulse.The duration of optional excitation light pulse
And/or intensity so that the maximum intensity of chemiluminescence emission light for example can be excited more than first caused by the tenth excitation pulse
The 50% of the maximum intensity of chemiluminescence emission light caused by pulse.The maximum intensity launched caused by tenth excitation pulse is very
75% of the maximum intensity of transmitting caused by the first excitation pulse can be extremely more than.Excitation light pulse can have substantially similar strong
Degree and duration.
Excitation wavelength is for example substantially equal to 680nm.Optional LED4 causes the peak wavelength and excitation wavelength of LED4
Matching, to provide maximal efficiency.Optional LED4 causes the peak wavelength of LED4 for example in the range of 675nm to 685nm.
Especially, the peak wavelength of LED4 is for example substantially equal to 680nm.
The light received from sample 2 may include such as launching light 11, reflected light and/or scattered light.The spectrum of receiving light point
The wavelength of amount can be in spectral detection band or outside spectral detection band.Can be measured from sample 2 in spectrum selectivity mode
Launching light 11 so that the spectral components outside detection band are not transferred, to detector 20.Can be surveyed in spectrum selectivity mode
Amount launching light 11 so that the spectral components outside detection band are not detected by detector 20.Especially, optional detection band is caused
The wavelength of exciting light（For example, 680nm）Outside detection band.The detector 20 of device 1 can be arranged to Detection wavelength and be in
Those spectral components of receiving light in detection band, and device 1 can be arranged to and prevent wavelength connecing outside the detection band
Receive the propagation of those spectral components of light.Spectral detection band is the wavelength model that the first cutoff wavelength and the second cutoff wavelength are limited
Enclose.First cutoff wavelength of detection band for example can be longer than or equal to 520nm, and the second cutoff wavelength of detection band for example can be short
In or equal to 620nm.Optional detection band enables detector 20 to detect chemiluminescence emission light 11, and allows to base
Propagation of this prevention exciting light 7 to detector 20.Device 1 may include one or more optical electivities point for limiting detection band
Amount.For example detection band can be limited by using dichroic beam splitters 8 and/or by using one or more wave filters.
The method can be that wherein donor bead and acceptor bead are by can specifically react to each other based on the neighbouring measure of pearl
And/or be connected with one or more biological analyte materials of pearl reaction.When using 670-690nm wavelength（For example with 680nm ripples
It is long）Red laser excited donor pearl when, periphery oxygen can be converted into excited singlet state by it.The creating singlet oxygen of release can spread
The sample well matrix of surrounding.If donor bead is connected with acceptor bead via particular organisms reaction to be checked, acceptor bead can be sufficient
It is enough close, to be excited by singlet state, and light transmitting can be produced.The wavelength of the light of transmitting for example can be in the range of 520-620nm.
Wavelength in the range of 520-620nm can correspond to such as blue-green, green or gold-tinted.The transmitting of light can with interaction level into than
Example.By measuring launching light, it is possible to establish significant degree that chemical reaction occurred and/or just in what property of determination sample
Matter.
Can take some time from creating singlet oxygen to the transmission of the energy of acceptor bead, and when the end and survey of excitation light pulse
When amount is postponed by existence time between chemiluminescence caused by the excitation light pulse, the signal to noise ratio of measurement can be improved.More precisely
Say that the time delay may imply that the end of excitation light pulse and measurement by chemiluminescence caused by the excitation light pulse in ground
Time period between the beginning of transmitting detection time section used.The length of time delay can for example be more than 1ms.Time delay
Length is for example substantially equal to 50ms.Pass by after predetermined time period from the transmission of excitation light pulse, can carry out
The measurement of launching light 11, wherein predetermined time period can for example be longer than 1ms.
By using the excitation wavelength of the second cutoff wavelength for being longer than detection band, conventional fluorescent can be substantially reduced or eliminated
Caused jamming pattern.The measurement of chemiluminescent light can be separated spectrally with excitation light pulse, so as to reduce background.Chemistry
The fluorescence that the measurement of luminous light temporarily can launch with sample 2 and/or well 3 is separated up to such as 50ms.When measurement creating singlet oxygen
During caused chemiluminescence emission, excitation wavelength can be longer than the second cutoff wavelength of detection band.The wavelength of background fluorescent emission can
It is longer than excitation wavelength.Therefore, be alternatively used for measure chemiluminescence emission detection band so that detection band spectrally not with it is glimmering
Light transmitting is overlapped.
The instantaneous signal obtained from emission detector 20 can with the instantaneous strength of chemiluminescence emission light 11 substantially into than
Example.The integration of the instantaneous signal obtained from emission detector 20 after single excitation light pulse can with the single exciting light arteries and veins
Integration after punching during integration time period from the intensity of the chemiluminescence emission light 11 of electromagnetic radiation is substantially proportional.Device
1 can be arranged to the integration for providing the instantaneous signal obtained from emission detector.Integration may indicate that AlphaScreen signals.Can
Selection of land, can standardize the integration by using the energy of excitation light pulse or intensity.Alternatively, the integration can divided by integration when
Between section length, to provide mean value.Alternatively, two or more integrated values can be with summed or be averaging, to improve
Certainty of measurement.
By using transmitting reference light source and scattering surface adjustable emission detector 20.Scattering surface can be arranged to temporarily
When ground to the non-reflective reference light source of emission detector 20 light.Transmitting inspection can be alternatively coupled to from the light of scattering surface reflection
Device 20 is surveyed, to calibrate the response of emission detector 20.Scattering surface can be for example white scattering surface or grey chromatic dispersion table
Face.Detector 20 can calibrate small-signal by using grey scattering surface.By using two or more with different cloudy
The scattering surface of shadow, the dynamic range of emission detector 20 can be calibrated.
Example 1
Using the AlphaScreen phosphotyrosines from Perkin Elmer（PT66）Detection kit（Production code member
6760602M）Carry out the long-time kinetic measurement of AlphaScreen signals.
In the measure, AlphaScreen donor beads and acceptor bead are combined together with biotinylated phosphoeptide.This shape
Into AlphaScreen compounds, apart from close, it is generated wherein donor bead and acceptor bead via creating singlet oxygen mechanism
AlphaScreen signals.Therefore, the AlphaScreen signals for measuring in the measure are proportional to biotinylated LCK-P peptides
Amount.
Perkin Elmer utilize 384 well Alphaplate using 25 l cumulative volumes（Production code member 6005359）Carry out dynamic
The test that mechanics AlphaScreen is determined.According to the instruction of manufacturer, mixing donor bead and acceptor bead.Then, 15 l pearls are mixed
The biotinylated LCK-P peptides of compound and 10 l variable concentrations are added to minitype plate well to reach 0.5,1.5,5,15 and
The ultimate density of 50fmol/ wells.Using eight repetition well measurements peptide dilutions, and buffering blank includes 16 repetitions.
Incubate assay plate in dark at room temperature one hour, measure the plate at room temperature using instrument of the invention afterwards.Measurement
It is set to：Excitation wavelength 680nm, launch wavelength 570nm, firing time 300ms, time delay 40ms and the time of integration
300ms.In the total time of 22 hours, one-shot measurement is carried out with dynamics form each two hour.
Fig. 5 A show the result of this long-time kinetic measurement that AlphaScreen signals are carried out with biotinylation peptide of phosphorylation.
Example 2
Using determining with identical in example 1, the short-time dynamics stability of AlphaScreen measurements is tested.With
AlphaScreen pearls（8 repetitions）Incubate the biotinylation peptide of phosphorylation of 0.5 fmol/ wells two minutes, and using same as above
Instrument is arranged and utilizes apparatus measures AlphaScreen signal of the invention.The measurement 20 times were repeated with six seconds intervals.
Fig. 5 B show the result using this short-time dynamics measurement of the AlphaScreen signals of biotinylation peptide of phosphorylation.
Example 3
Omnidirectional pearl（Omnibead）It is the spy of the checking and the calibration that have been designed for the instrument used in AlphaScreen technologies
The AlphaScreen pearls of different type（Perkin Elmer, production code member 6760626M）.Omnidirectional pearl is only strong containing generating
The pearl of the single type of AlphaScreen signals.
In the measure, with 10 mM PBSs dilution omnidirectional pearl, with eight repetitions by the every of the l of pH 7.2 and 25
Individual dilution is added in 384- well Alphaplate.Kinetic measurement is immediately begun to after liquid relief, and is utilized at room temperature
The instrument of the present invention measured an assay plate in 12 hours per 30 minutes.Measure setup is as above：Excitation wavelength 680nm, transmitting
Wavelength 570nm, firing time 300ms, time delay 40ms and time of integration 300ms.
Fig. 5 C show the knot of this long-time kinetic measurement that AlphaScreen signals are carried out with omnidirectional pearl correction pearl
Really.
Can be seen that in TRF measurement sequences from Fig. 5 A-5C, the result of exemplary measurement keeps fairly constant.
Accompanying drawing and it is described above shown in specific exemplary embodiment of the invention be not necessarily to be construed as limit.Ability
Field technique personnel can be with many obvious ways these embodiments of modifications and changes within the scope of the appended claims.Thus,
The present invention is not limited to embodiments described above.
Claims (23)
1. it is a kind of to be used to be placed on sample well（3）In fluid sample（2）Optical measurement method, wherein in the sample well
Sample be exposed to from excitation source（4）The exciting light of first wave strong point（7）, donor of the exciting light from fluid sample point
Son generates the set of singlet state molecular oxygen, and the singlet state molecular oxygen causes described receiving with the acceptor molecule reaction in fluid sample
Body molecule launches chemiluminescence emission light with second wave length（11）, the second wave length is than the first wave length, and wherein uses
The launching light that detectors measure is produced by the exciting light, it is characterised in that the excitation source is LED（4）.
2. method according to claim 1, wherein from LED（4）Exciting light（7）It is focused on sample well（3）Open
Mouth region domain, and use confocal arrangement（9）Launching light is collected from the open area of the sample well（11）.
3. method according to claim 1 and 2, the wherein exciting light（7）It is imported into the sample well（3）And from confocal cloth
Put（9）In sample well collect launching light（11）, the wherein excitation source（4）Image（15）It is focused on opening for the sample well
On at the top of mouthful, and the optical receiving region of the detector（14）Image（16）It is focused in the open top of the sample well.
4. method according to claim 3, the wherein excitation source（4）Described image（15）Less than the sample well（3）
Opening.
5. method according to claim 3, the wherein optical receiving region of detector（14）Described image（16）It is more than
The sample well（3）Opening.
6. method according to any one of claim 1 to 5, wherein to sample（2）Send excitation light pulse, and from swash
The transmission of LED pulse is pass by after predetermined time period, carries out launching light（11）Measurement.
7. method according to claim 6, wherein the predetermined time period is longer than 1ms.
8. method according to any one of claim 1 to 7, wherein preferably by calibration process calibration separately and independently
The exciting light and launching light.
9. method according to any one of claim 1 to 8, including：
The a part of excitation light pulse of-coupling is to exciting reference detector（19）, and
- based on exciting reference detector（19）Output be adjusted to LED（4）Electric current so that carry out self-excitation reference detector
（19）Output signal be retained as during the excitation light pulse it is substantially invariable.
10. method according to claim 9, including by using pi controller to LED（4）Electric current is provided.
11. methods according to any one of claim 1 to 10, the wherein LED（4）In 680nm emitted at wavelengths light.
12. methods according to any one of claim 1 to 11, the wherein LED（4）Peak wavelength arrive in 675nm
In the range of 685nm.
13. methods according to any one of claim 1 to 12, wherein measuring the launching light in spectrum selectivity mode
（11）So that the spectral components outside detection band are not transferred, to the detector（20）, excitation source（4）First wave length
（680nm）Outside detection band, device（1）The first cutoff wavelength of detection band be longer than or equal to 520nm, and detection band
Second cutoff wavelength is shorter than or equal to 620nm.
14. methods according to any one of claim 1 to 13, the wherein sample are excited, and corresponding transmitting quilt
Read at least twice.
15. methods according to any one of claim 1 to 14, including：
- irradiate the sample with the first excitation light pulse（2）, so as to from sample（2）Donor molecule generate singlet state molecular oxygen
First set,
- by the first set from singlet state molecular oxygen to sample（2）Acceptor molecule transmission energy cause the first chemiluminescence
Launching light（11）Transmitting,
- measure the first chemiluminescence emission light（11）,
- irradiate the sample with the second excitation light pulse（2）, to generate the second of singlet state molecular oxygen from the donor molecule of sample
Set,
- by the second set from singlet state molecular oxygen to sample（2）Acceptor molecule transmission energy cause the second chemiluminescence
Launching light（11）Transmitting, and
- measure the second chemiluminescence emission light（11）,
The wherein sample（2）By the first excitation light pulse and by the irradiation of the second excitation light pulse so that second chemiluminescence is sent out
The maximum intensity of light is penetrated more than or equal to the 80% of the maximum intensity of the first chemiluminescence emission light, and wherein this first swashs
Time delay between the end of LED pulse and the second excitation light pulse is more than 30ms.
16. methods according to any one of claim 1 to 15, including being arranged using lens（9）By focusing on the LED
（4）Light providing narrow beam so that form the LED in the focal point of the narrow beam（4）Focusedimage（15）, wherein
The lens arrangement（9）Numerical aperture（NA）More than 0.20.
17. one kind are used to be placed on sample well（3）In fluid sample（2）Optical measurement device（1）, the device include use
In in first wave strong point to exciting light（7）The excitation source of the sample in exposed sample well（4）, and at second wave length
From sample detection and measure chemiluminescence emission light（11）Detector, the second wave length be less than the first wave length, its feature
It is that the excitation source is LED（4）.
18. devices according to claim 17（1）, the wherein device（1）Including for focusing on sample well（3）Opening
Excitation source in region（4）Image（15）Focus set（9）, and in the open area for focusing on sample well
The optical receiving region of detector（14）Image（16）Equipment（9）.
19. devices according to claim 17 or 18（1）, the wherein excitation source（4）For pulse excitation light source.
20. devices according to any one of claim 17 to 19（1）, wherein the device include excite optical filter（6）、
Transmitting optical filter（12）, excite and launch beam splitter（8）And sample well（3）Spuious light shield above（10）.
21. devices according to any one of claim 17 to 20（1）, wherein the device include excite beam splitter（18）
With for calibrating exciting light（7）Excite reference detector（19）, and/or transmitting reference light source（21）And for calibrating transmitting
The white reflective scattering surface of detector.
22. devices according to any one of claim 17 to 21（1）, the wherein LED（4）For the LED at 680nm wavelength
Launching light.
23. devices according to any one of claim 17 to 22（1）, the wherein detector is photomultiplier（20）.
Applications Claiming Priority (5)
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| FI20145774 | 2014-09-05 | ||
| FI20145774 | 2014-09-05 | ||
| FI20146110 | 2014-12-17 | ||
| FI20146110 | 2014-12-17 | ||
| PCT/FI2015/050578 WO2016034775A1 (en) | 2014-09-05 | 2015-09-03 | Method and apparatus for optical measurement of liquid sample |
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| CN106662530A true CN106662530A (en) | 2017-05-10 |
Family
ID=55439171
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|---|---|
| US (1) | US10677734B2 (en) |
| EP (1) | EP3189325B1 (en) |
| CN (1) | CN106662530A (en) |
| WO (1) | WO2016034775A1 (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113711013A (en) * | 2019-02-04 | 2021-11-26 | 生命科技控股私人有限公司 | Sample analysis method, analysis device, and computer program |
Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20200025685A1 (en) * | 2016-12-15 | 2020-01-23 | Codiak Biosciences, Inc. | Methods of measuring exosomes using intrinsic fluorescence |
| US11137343B2 (en) * | 2019-12-10 | 2021-10-05 | Trustees Of Boston University | Apparatus and method for biomolecular analysis |
Citations (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH11311603A (en) * | 1998-04-30 | 1999-11-09 | Jasco Corp | Measuring apparatus |
| WO2000050860A1 (en) * | 1999-02-26 | 2000-08-31 | Packard Instrument Company, Inc. | Producing and measuring light and determining the amounts of analytes in microplate wells |
| CN1301357A (en) * | 1998-03-16 | 2001-06-27 | 普雷勒克斯公司 | Confocal microscopy imaging system |
| CN1771429A (en) * | 2003-02-27 | 2006-05-10 | 美国瓦里安澳大利亚有限公司 | Spectrophotometer |
| WO2009056669A1 (en) * | 2007-10-31 | 2009-05-07 | Wallac Oy | Sample measurement system |
| CN101514962A (en) * | 2009-03-31 | 2009-08-26 | 东北电力大学 | Fluorescent mark immunity test strip detector |
| CN102292924A (en) * | 2009-01-26 | 2011-12-21 | 沃拉克有限公司 | An optical measurement instrument with data transmission |
| CN102714904A (en) * | 2009-10-14 | 2012-10-03 | 惠普发展公司，有限责任合伙企业 | Stabilized light source having luminance feedback control |
| WO2013163129A1 (en) * | 2012-04-23 | 2013-10-31 | Siemens Healthcare Diagnostics Inc. | Biological assay sample analyzer |
Family Cites Families (21)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6251581B1 (en) | 1991-05-22 | 2001-06-26 | Dade Behring Marburg Gmbh | Assay method utilizing induced luminescence |
| US6097025A (en) * | 1997-10-31 | 2000-08-01 | Ljl Biosystems, Inc. | Light detection device having an optical-path switching mechanism |
| US6496260B1 (en) * | 1998-12-23 | 2002-12-17 | Molecular Devices Corp. | Vertical-beam photometer for determination of light absorption pathlength |
| JP3597729B2 (en) * | 1999-05-11 | 2004-12-08 | 日立ソフトウエアエンジニアリング株式会社 | Fluorescence metering method and fluorescence meter |
| US7423750B2 (en) * | 2001-11-29 | 2008-09-09 | Applera Corporation | Configurations, systems, and methods for optical scanning with at least one first relative angular motion and at least one second angular motion or at least one linear motion |
| WO2001069302A2 (en) * | 2000-03-13 | 2001-09-20 | Genospectra, Inc. | Fiber optic scanner |
| US6545758B1 (en) | 2000-08-17 | 2003-04-08 | Perry Sandstrom | Microarray detector and synthesizer |
| EP1436596A2 (en) * | 2001-09-28 | 2004-07-14 | Ciencia, Incorporated | Compact multiwavelength phase fluorometer |
| DE10202050A1 (en) * | 2002-01-18 | 2003-07-31 | Siemens Ag | Imaging of live small animals using luminescence techniques for biological, medical and pharmaceutical research, whereby LEDs are used as a cost effective and low-power light source that provides sufficient excitation light energy |
| FI20030867A (en) | 2003-06-10 | 2004-12-11 | Wallac Oy | Optical measuring method and laboratory measuring apparatus |
| FI20031357A0 (en) | 2003-09-19 | 2003-09-19 | Wallac Oy | Multipurpose apparatus for optical measurement of samples |
| US20070247628A1 (en) | 2004-09-10 | 2007-10-25 | Wallac Oy | Instrumentation and Method Adapted For Optical Measurement of an Amplified Luminescent Proximity Homogeneous Assay |
| JP4660691B2 (en) * | 2005-04-08 | 2011-03-30 | コニカミノルタセンシング株式会社 | Optical property measuring method of fluorescent sample and optical property measuring apparatus using the same |
| US8496879B2 (en) * | 2006-02-08 | 2013-07-30 | Molecular Devices, Llc | Optical detection utilizing cartridge with tunable filter assembly |
| US7700928B2 (en) * | 2007-01-25 | 2010-04-20 | Etaluma, Inc. | Apparatus and method for interleaving detection of fluorescence and luminescence |
| FI20070171A0 (en) | 2007-02-28 | 2007-02-28 | Hidex Oy | Bioassay and bioassay equipment |
| FI20075439A0 (en) | 2007-06-12 | 2007-06-12 | Wallac Oy | Automated instrumentation and method for measuring samples |
| CN202126403U (en) | 2011-06-05 | 2012-01-25 | 陈国颖 | Antibiotics analysis device based on resolution of excitation time characteristics of fluorescence spectrum |
| DE102013207479B3 (en) | 2013-04-24 | 2014-10-02 | Bundesdruckerei Gmbh | Method for the rapid determination of the absolute luminescence intensity |
| US11536716B2 (en) * | 2015-06-25 | 2022-12-27 | Hoffmann-La Roche Inc. | Cell based assay for determining antibody or ligand binding and function |
| WO2018052481A1 (en) * | 2016-09-13 | 2018-03-22 | Insight Instruments, Inc. | Homogenous and heterogeneous assays and systems for determination of ocular biomarkers |
-
2015
- 2015-09-03 CN CN201580047404.9A patent/CN106662530A/en active Pending
- 2015-09-03 EP EP15838441.2A patent/EP3189325B1/en active Active
- 2015-09-03 US US15/505,486 patent/US10677734B2/en active Active
- 2015-09-03 WO PCT/FI2015/050578 patent/WO2016034775A1/en active Application Filing
Patent Citations (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1301357A (en) * | 1998-03-16 | 2001-06-27 | 普雷勒克斯公司 | Confocal microscopy imaging system |
| JPH11311603A (en) * | 1998-04-30 | 1999-11-09 | Jasco Corp | Measuring apparatus |
| WO2000050860A1 (en) * | 1999-02-26 | 2000-08-31 | Packard Instrument Company, Inc. | Producing and measuring light and determining the amounts of analytes in microplate wells |
| CN1771429A (en) * | 2003-02-27 | 2006-05-10 | 美国瓦里安澳大利亚有限公司 | Spectrophotometer |
| WO2009056669A1 (en) * | 2007-10-31 | 2009-05-07 | Wallac Oy | Sample measurement system |
| CN102292924A (en) * | 2009-01-26 | 2011-12-21 | 沃拉克有限公司 | An optical measurement instrument with data transmission |
| CN101514962A (en) * | 2009-03-31 | 2009-08-26 | 东北电力大学 | Fluorescent mark immunity test strip detector |
| CN102714904A (en) * | 2009-10-14 | 2012-10-03 | 惠普发展公司，有限责任合伙企业 | Stabilized light source having luminance feedback control |
| WO2013163129A1 (en) * | 2012-04-23 | 2013-10-31 | Siemens Healthcare Diagnostics Inc. | Biological assay sample analyzer |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113711013A (en) * | 2019-02-04 | 2021-11-26 | 生命科技控股私人有限公司 | Sample analysis method, analysis device, and computer program |
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| Publication number | Publication date |
|---|---|
| US10677734B2 (en) | 2020-06-09 |
| EP3189325A1 (en) | 2017-07-12 |
| WO2016034775A1 (en) | 2016-03-10 |
| EP3189325B1 (en) | 2022-02-09 |
| US20170254756A1 (en) | 2017-09-07 |
| EP3189325A4 (en) | 2018-06-06 |
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